Key Product Details

Validated by

Independent Antibodies, Biological Validation

Species Reactivity

Validated:

Mouse

Cited:

Mouse

Applications

Validated:

Western Blot, Immunoprecipitation

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all IRF3 Primary Antibodies »

Product Specifications

Immunogen

The immunogen for this product maps to a region between residue 375 and 419 of mouse Interferon Regulatory Factor 3 using the numbering given in entry NP_058545.1 (GeneID 54131).

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

47 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for IRF3 Antibody - BSA Free

Western Blot: IRF3 Antibody [NBP1-78769]

Western Blot: IRF3 Antibody [NBP1-78769]

Western Blot: IRF3 Antibody [NBP1-78769] - Whole cell lysate from mouse bone marrow dendritic (D; 50 and 15 mcg for WB; 1 mg for IP, 20% of IP loaded), and NIH3T3 (M; 50 mcg) cells. Affinity purified rabbit anti-IRF3 antibody used for WB at 0.4 mcg/ml (A) and 1 mcg/ml (B) and used for IP at 6 mcg/mg lysate. IRF3 was also immunoprecipitated by rabbit anti-IRF3 antibody NBP1-78768 which recognizes a disparate epitope.
IRF3 Antibody - BSA Free

Western Blot: IRF3 Antibody - BSA Free [NBP1-78769] -

Auranofin activates the cellular autophagy pathway to degrade IRF3 protein.A, HT1080 cells, pretreated with auranofin (AF), were transfected with polyI:C (pIC+LF) for 24 h, when LC3 and IRF3 were analyzed by immunoblot. B and C, MDA-MB-453 cells were treated with AF for the indicated times when the p62 levels were analyzed by immunoblot (B), and the immunoblots from the biological replicates were quantified using ImageJ (C). D, RAW264.7 cells were treated with AF for the indicated times when the p62 levels were analyzed by immunoblot. E–G, HT1080 cells, stably expressing ATG5-specific or nontargeting (NT) shRNA, were treated with AF for the indicated times, when the levels of p62 (E and F) or IRF3 (G) were analyzed by immunoblot. The quantification in F is from the biological replicates. ATG5 knockdown levels were analyzed by immunoblot (E, lower panel). H, HT1080 cells, stably expressing ATG5-specific or nontargeting (NT) shRNA, were treated with AF for the indicated times, when the levels of RIG-I, MAVS, TRAF2, TBK1, BAX, and ATG5 were analyzed by immunoblot. Veh, Vehicle (DMSO). Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S0021925821010772), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF3 Antibody - BSA Free

Western Blot: IRF3 Antibody - BSA Free [NBP1-78769] -

Auranofin promotes the degradation of IRF3 protein.A, HT1080 cells, pretreated with auranofin (AF), were transfected with poly(I:C) (pIC+LF) and the phosphorylated (Ser396) and total IRF3 were analyzed by immunoblot after 8 h. B, MDA-MB-453 cells, pretreated with AF, were transfected with polyI:C (pIC+LF) for the indicated times when the IRF3 levels were analyzed by immunoblot. C and D, MDA-MB-453 cells were treated with AF for the indicated times when the IRF3 levels were analyzed by immunoblot (C), and the immunoblots from the biological replicates were quantified using ImageJ (D). E, HT1080 cells were treated with AF for the indicated times when the IRF3 protein levels were analyzed by immunoblot. F, RAW264.7 cells were treated with AF at the indicated concentrations for 16 h, when the levels of Irf3 were analyzed by immunoblot. G, RAW264.7 cells were treated with AF (5 μM) for the indicated times when the levels of Irf3 were analyzed by immunoblot. H, HT1080 cells were treated with AF for the indicated times, and the BAX protein levels were analyzed by immunoblot. I, HT1080 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of AF and analyzed for phosphorylated (on Ser536) and total p65 by immunoblot after 4 h. J, HT1080 cells were treated with interferon (hIFN beta, 1000 U/ml) in the absence or the presence of AF and analyzed for phosphorylated (on Ser727) and total STAT1 by immunoblot after 2 h. K, HT1080 cells were treated with AF at the indicated concentrations for 8 h when cell viability was assessed by trypan blue exclusion assay. Veh, Vehicle (DMSO). Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S0021925821010772), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF3 Antibody - BSA Free

Western Blot: IRF3 Antibody - BSA Free [NBP1-78769] -

Autophagic degradation inhibits the transcriptional activity of IRF3.A, MDA-MB-453 cells were treated with Hank's balanced salt solution (HBSS) for the indicated times when the protein levels of IRF3 were analyzed by immunoblot. B and C, MDA-MB-453 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of HBSS for the indicated times, when the protein levels of pIRF3 and IRF3 (B) and IFIT3 (C) were analyzed by immunoblot. D–F, MDA-MB-453 (D, E) or RAW264.7 (F) cells were transfected with polyI:C (pIC+LF) in the absence or the presence of HBSS for 8 h, when IFIT3 (D), IFNB1 (E), and Ifit1 (F) mRNA levels were analyzed by qRT-PCR. G, HT1080 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of HBSS and phosphorylated (on Ser172), and total TBK1 was analyzed by immunoblot after 4 h. H, HT1080 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of HBSS and phosphorylated (on Thr180/Tyr182) and total p38 MAPK was analyzed by immunoblot after 4 h. I, U4C (Wt) and IRF3-overexpressing U4C (IRF3hi) cells were transfected with polyI:C (pIC+LF) in the absence or the presence of HBSS, and IFIT3 and IRF3 were analyzed by immunoblot at the indicated times posttransfection. ∗ indicates p < 0.05. Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S0021925821010772), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF3 Antibody - BSA Free

Western Blot: IRF3 Antibody - BSA Free [NBP1-78769] -

Autophagic degradation of IRF3 by auranofin inhibits palmitic-acid-induced apoptotic cell death in human hepatocytes.A, Huh7 cells were treated with palmitic acid (PA) or vehicle (bovine serum albumin, BSA) for 24 h, when the caspase-3 activity was measured. B and C, Huh7 cells were treated with PA, and apoptotic cell death was analyzed by immunoblot of cleaved PARP (B) or bright field microscopy (C). D and E, Huh7 cells were treated with PA, and the caspase activity (D), and cleaved PARP (E) were analyzed at the indicated times post-treatment. F and G, Huh7 cells were pretreated with auranofin (AF) at the indicated concentrations and treated with PA. Caspase activity (F) and cleaved PARP (G) were measured 16 h posttreatment. H, Huh7 cells were treated with AF at the indicated concentrations for 8 h when cell viability was assessed by trypan blue exclusion assay. I, Huh7 cells were transfected with nontargeting (NT) or IRF3-specific siRNA, and the cells were analyzed for caspase-3 activity upon PA treatment for 16 h. The lower panel indicates the knockdown levels of IRF3 protein. J and K, Huh7 cells were pretreated with AF at the indicated concentrations and treated with PA. IRF3 (J) and p62 (K) were analyzed 16 h posttreatment. L, Huh7 cells were treated with AF, as indicated, and the LC3 levels were analyzed by immunoblot. Veh, Vehicle (DMSO), ∗ indicates p < 0.05, NS, nonsignificant, scale bar, 100 μm. Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S0021925821010772), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF3 Antibody - BSA Free

Western Blot: IRF3 Antibody - BSA Free [NBP1-78769] -

Auranofin promotes the degradation of IRF3 protein.A, HT1080 cells, pretreated with auranofin (AF), were transfected with poly(I:C) (pIC+LF) and the phosphorylated (Ser396) and total IRF3 were analyzed by immunoblot after 8 h. B, MDA-MB-453 cells, pretreated with AF, were transfected with polyI:C (pIC+LF) for the indicated times when the IRF3 levels were analyzed by immunoblot. C and D, MDA-MB-453 cells were treated with AF for the indicated times when the IRF3 levels were analyzed by immunoblot (C), and the immunoblots from the biological replicates were quantified using ImageJ (D). E, HT1080 cells were treated with AF for the indicated times when the IRF3 protein levels were analyzed by immunoblot. F, RAW264.7 cells were treated with AF at the indicated concentrations for 16 h, when the levels of Irf3 were analyzed by immunoblot. G, RAW264.7 cells were treated with AF (5 μM) for the indicated times when the levels of Irf3 were analyzed by immunoblot. H, HT1080 cells were treated with AF for the indicated times, and the BAX protein levels were analyzed by immunoblot. I, HT1080 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of AF and analyzed for phosphorylated (on Ser536) and total p65 by immunoblot after 4 h. J, HT1080 cells were treated with interferon (hIFN beta, 1000 U/ml) in the absence or the presence of AF and analyzed for phosphorylated (on Ser727) and total STAT1 by immunoblot after 2 h. K, HT1080 cells were treated with AF at the indicated concentrations for 8 h when cell viability was assessed by trypan blue exclusion assay. Veh, Vehicle (DMSO). Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S0021925821010772), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF3 Antibody - BSA Free

Western Blot: IRF3 Antibody - BSA Free [NBP1-78769] -

Auranofin promotes the degradation of IRF3 protein.A, HT1080 cells, pretreated with auranofin (AF), were transfected with poly(I:C) (pIC+LF) and the phosphorylated (Ser396) and total IRF3 were analyzed by immunoblot after 8 h. B, MDA-MB-453 cells, pretreated with AF, were transfected with polyI:C (pIC+LF) for the indicated times when the IRF3 levels were analyzed by immunoblot. C and D, MDA-MB-453 cells were treated with AF for the indicated times when the IRF3 levels were analyzed by immunoblot (C), and the immunoblots from the biological replicates were quantified using ImageJ (D). E, HT1080 cells were treated with AF for the indicated times when the IRF3 protein levels were analyzed by immunoblot. F, RAW264.7 cells were treated with AF at the indicated concentrations for 16 h, when the levels of Irf3 were analyzed by immunoblot. G, RAW264.7 cells were treated with AF (5 μM) for the indicated times when the levels of Irf3 were analyzed by immunoblot. H, HT1080 cells were treated with AF for the indicated times, and the BAX protein levels were analyzed by immunoblot. I, HT1080 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of AF and analyzed for phosphorylated (on Ser536) and total p65 by immunoblot after 4 h. J, HT1080 cells were treated with interferon (hIFN beta, 1000 U/ml) in the absence or the presence of AF and analyzed for phosphorylated (on Ser727) and total STAT1 by immunoblot after 2 h. K, HT1080 cells were treated with AF at the indicated concentrations for 8 h when cell viability was assessed by trypan blue exclusion assay. Veh, Vehicle (DMSO). Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S0021925821010772), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF3 Antibody - BSA Free

Western Blot: IRF3 Antibody - BSA Free [NBP1-78769] -

Auranofin promotes the degradation of IRF3 protein.A, HT1080 cells, pretreated with auranofin (AF), were transfected with poly(I:C) (pIC+LF) and the phosphorylated (Ser396) and total IRF3 were analyzed by immunoblot after 8 h. B, MDA-MB-453 cells, pretreated with AF, were transfected with polyI:C (pIC+LF) for the indicated times when the IRF3 levels were analyzed by immunoblot. C and D, MDA-MB-453 cells were treated with AF for the indicated times when the IRF3 levels were analyzed by immunoblot (C), and the immunoblots from the biological replicates were quantified using ImageJ (D). E, HT1080 cells were treated with AF for the indicated times when the IRF3 protein levels were analyzed by immunoblot. F, RAW264.7 cells were treated with AF at the indicated concentrations for 16 h, when the levels of Irf3 were analyzed by immunoblot. G, RAW264.7 cells were treated with AF (5 μM) for the indicated times when the levels of Irf3 were analyzed by immunoblot. H, HT1080 cells were treated with AF for the indicated times, and the BAX protein levels were analyzed by immunoblot. I, HT1080 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of AF and analyzed for phosphorylated (on Ser536) and total p65 by immunoblot after 4 h. J, HT1080 cells were treated with interferon (hIFN beta, 1000 U/ml) in the absence or the presence of AF and analyzed for phosphorylated (on Ser727) and total STAT1 by immunoblot after 2 h. K, HT1080 cells were treated with AF at the indicated concentrations for 8 h when cell viability was assessed by trypan blue exclusion assay. Veh, Vehicle (DMSO). Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S0021925821010772), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF3 Antibody - BSA Free

Western Blot: IRF3 Antibody - BSA Free [NBP1-78769] -

Auranofin inhibits the proapoptotic activity of IRF3.A, HT1080 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of auranofin (AF), and the culture fields were analyzed by bright field microscopy 8 h post-pIC stimulation. B, HT1080 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of AF, at the indicated concentrations, for 16 h, when cleaved PARP was analyzed by immunoblot. C, HT1080 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of AF for the indicated times when the caspase-3 activity was measured. D, MDA-MB-453 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of AF (at the indicated concentrations) for 16 h, when cleaved PARP was analyzed by immunoblot. E, MDA-MB-453 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of AF for the indicated times when the caspase-3 activity was measured. F, HT1080 cells were transfected with poly(dA:dT) in the absence or the presence of AF for 16 h, when the cleaved PARP was analyzed by immunoblot. G, HT1080 cells were transfected with poly(dA:dT) in the absence or the presence of AF for 16 h, when the caspase-3 activity was measured. H, Wt and IRF3−/− HT1080 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of AF for 8 h when the caspase-3 activity was measured. IRF3 protein expression is shown by immunoblot in the inset. Veh, Vehicle (DMSO), ∗ indicates p < 0.05, NS, nonsignificant, scale bar, 100 μm. Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S0021925821010772), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF3 Antibody - BSA Free

Western Blot: IRF3 Antibody - BSA Free [NBP1-78769] -

Auranofin promotes the degradation of IRF3 protein.A, HT1080 cells, pretreated with auranofin (AF), were transfected with poly(I:C) (pIC+LF) and the phosphorylated (Ser396) and total IRF3 were analyzed by immunoblot after 8 h. B, MDA-MB-453 cells, pretreated with AF, were transfected with polyI:C (pIC+LF) for the indicated times when the IRF3 levels were analyzed by immunoblot. C and D, MDA-MB-453 cells were treated with AF for the indicated times when the IRF3 levels were analyzed by immunoblot (C), and the immunoblots from the biological replicates were quantified using ImageJ (D). E, HT1080 cells were treated with AF for the indicated times when the IRF3 protein levels were analyzed by immunoblot. F, RAW264.7 cells were treated with AF at the indicated concentrations for 16 h, when the levels of Irf3 were analyzed by immunoblot. G, RAW264.7 cells were treated with AF (5 μM) for the indicated times when the levels of Irf3 were analyzed by immunoblot. H, HT1080 cells were treated with AF for the indicated times, and the BAX protein levels were analyzed by immunoblot. I, HT1080 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of AF and analyzed for phosphorylated (on Ser536) and total p65 by immunoblot after 4 h. J, HT1080 cells were treated with interferon (hIFN beta, 1000 U/ml) in the absence or the presence of AF and analyzed for phosphorylated (on Ser727) and total STAT1 by immunoblot after 2 h. K, HT1080 cells were treated with AF at the indicated concentrations for 8 h when cell viability was assessed by trypan blue exclusion assay. Veh, Vehicle (DMSO). Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S0021925821010772), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF3 Antibody - BSA Free

Western Blot: IRF3 Antibody - BSA Free [NBP1-78769] -

Auranofin activates the cellular autophagy pathway to degrade IRF3 protein.A, HT1080 cells, pretreated with auranofin (AF), were transfected with polyI:C (pIC+LF) for 24 h, when LC3 and IRF3 were analyzed by immunoblot. B and C, MDA-MB-453 cells were treated with AF for the indicated times when the p62 levels were analyzed by immunoblot (B), and the immunoblots from the biological replicates were quantified using ImageJ (C). D, RAW264.7 cells were treated with AF for the indicated times when the p62 levels were analyzed by immunoblot. E–G, HT1080 cells, stably expressing ATG5-specific or nontargeting (NT) shRNA, were treated with AF for the indicated times, when the levels of p62 (E and F) or IRF3 (G) were analyzed by immunoblot. The quantification in F is from the biological replicates. ATG5 knockdown levels were analyzed by immunoblot (E, lower panel). H, HT1080 cells, stably expressing ATG5-specific or nontargeting (NT) shRNA, were treated with AF for the indicated times, when the levels of RIG-I, MAVS, TRAF2, TBK1, BAX, and ATG5 were analyzed by immunoblot. Veh, Vehicle (DMSO). Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S0021925821010772), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF3 Antibody - BSA Free

Western Blot: IRF3 Antibody - BSA Free [NBP1-78769] -

Autophagic degradation inhibits the transcriptional activity of IRF3.A, MDA-MB-453 cells were treated with Hank's balanced salt solution (HBSS) for the indicated times when the protein levels of IRF3 were analyzed by immunoblot. B and C, MDA-MB-453 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of HBSS for the indicated times, when the protein levels of pIRF3 and IRF3 (B) and IFIT3 (C) were analyzed by immunoblot. D–F, MDA-MB-453 (D, E) or RAW264.7 (F) cells were transfected with polyI:C (pIC+LF) in the absence or the presence of HBSS for 8 h, when IFIT3 (D), IFNB1 (E), and Ifit1 (F) mRNA levels were analyzed by qRT-PCR. G, HT1080 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of HBSS and phosphorylated (on Ser172), and total TBK1 was analyzed by immunoblot after 4 h. H, HT1080 cells were transfected with polyI:C (pIC+LF) in the absence or the presence of HBSS and phosphorylated (on Thr180/Tyr182) and total p38 MAPK was analyzed by immunoblot after 4 h. I, U4C (Wt) and IRF3-overexpressing U4C (IRF3hi) cells were transfected with polyI:C (pIC+LF) in the absence or the presence of HBSS, and IFIT3 and IRF3 were analyzed by immunoblot at the indicated times posttransfection. ∗ indicates p < 0.05. Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S0021925821010772), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for IRF3 Antibody - BSA Free

Application
Recommended Usage

Immunoprecipitation

2-10 ug/mg lysate

Western Blot

1:1000-1:2500
Application Notes
Western blot of lysates performed using standard western blot reagents and 4-20% SDS-PAGE.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris-Citrate/Phosphate (pH 7.0 - 8.0)

Format

BSA Free

Preservative

0.09% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C. Do not freeze.

Background: IRF3

IRF3 is a family of interferon (IFN) regulatory factors (IRFs) have been shown to play a role in transcription of IFN genes as well as IFN-stimulated genes. The IRF-3 gene encodes a 50-kDa protein that binds specifically to the IFN-stimulated response element (ISRE) but not to the IRF-1 binding site PRD-I. Overexpression of IRF-3 stimulates expression of the IFN-stimulated gene 15 (ISG15) promoter, an ISRE-containing promoter. he high amino acid homology between IRF-3 and ISG factor 3 gamma polypeptide (ISGF3 gamma) and their similar binding properties indicate that, like ISGF3 gamma, IRF-3 may activate transcription by complex formation with other transcriptional factors, possibly members of the Stat family (1). IRF-3 is expressed constitutively in a variety of tissues, and the relative levels of IRF-3 mRNA do not change in virus-infected or IFN-treated cells. (2). In uninfected cells, the IRF-3 component of DRAF1 resides in the cytoplasm. The cytoplasmic localization of IRF-3 is dependent on a nuclear export signal, and IRF-3 is recognized by the chromosome region maintenance 1 (CRM1) (also known as exportin 1) shuttling receptor. Following infection and specific phosphorylation, IRF-3 accumulates in the nucleus where it associates with CBP and p300 (3).

Long Name

Interferon Regulatory Factor 3

Alternate Names

interferon regulatory factor 3, IRF-3

Entrez Gene IDs

54131 (Mouse)

Gene Symbol

IRF3

UniProt

P70671

Additional IRF3 Products

Product Documents for IRF3 Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for IRF3 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for IRF3 Antibody - BSA Free

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